Activation of a peroxisomal Pichia pastoris D-amino acid oxidase, which uses d-alanine as a preferred substrate, depends on pyruvate carboxylase.

Abstract:

:d-Amino acid oxidase (DAO) is an important flavo-enzyme that catalyzes the oxidative deamination of d-amino acids into the corresponding alpha-keto acid, ammonia and H(2)O(2). We identified two amino acid oxidases in the methylotrophic yeast Pichia pastoris: Dao1p, which preferentially uses d-alanine as a substrate, and Dao2p, which uses d-aspartate as a preferred substrate. Dao1p has a molecular mass of 38.2 kDa and a pH optimum of 9.6. This enzyme was localized to peroxisomes, albeit a typical peroxisomal targeting signal is lacking. Interestingly, P. pastoris mutant strains, defective in the enzyme pyruvate carboxylase, showed a pronounced growth defect on d-alanine, concomitant with a significant reduction in Dao1p activity relative to the wild-type control. This indicates that pyruvate carboxylase functions in import and/or activation of P. pastoris Dao1p.

journal_name

FEMS Yeast Res

journal_title

FEMS yeast research

authors

Klompmaker SH,Kilic A,Baerends RJ,Veenhuis M,van der Klei IJ

doi

10.1111/j.1567-1364.2010.00647.x

subject

Has Abstract

pub_date

2010-09-01 00:00:00

pages

708-16

issue

6

eissn

1567-1356

issn

1567-1364

pii

FYR647

journal_volume

10

pub_type

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